Two-hybrid system: yeast

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey was previously cloned reagent).

Two-hybrid system: yeast

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey was previously cloned reagent).

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey was previously cloned reagent).

Lhr binds to Su(var)205 (HP1) and HP6, but not to HP4, HP5 or rhi in this yeast two hybrid study.

The interaction of Lhr with Su(var)205 is conserved in other Drosophila species.

Two-hybrid system: yeast GAL4-BD/GAL4-AD

Su(var)205 was isolated in Hmr, Lhr and tandem Hmr-Lhr affinity purification experiments.

Source was cell extract of S2 cell line; bait produced from transfected construct; prey produced from endogenous gene.

Su(var)205 coimmunoprecipitates Hmr and Lhr, but not HP6.

Source was nuclear extract of S2 cell line; bait produced from transfected construct; prey produced from endogenous gene.

Source was cross-linked nuclear extract of S2 cell line; bait produced from transfected construct; prey produced from endogenous gene.

Crude nuclear extracts were cross-linked using formaldehyde, sonicated, and subjected to ProteinA affinity purification with rabbit IgG agarose beads eluted under denaturing conditions, and subsequently using streptavidin agarose beads for Bio affinity purification. Purified peptides were eluted by on-bead trypsin digestion and mass spectrometric analysis.

Enrichment of prey protein was compared to input, mock and unrelated BioTAP affinity purifications using a statistical method (Bamse) developed by the authors to control for multiple sources of bias.